JP5615484B2 - 5α-reductase inhibitor - Google Patents

Description

  The present invention relates to a 5α-reductase inhibitor, a food / beverage product imparted with a 5α-reductase inhibitory function, and a pharmaceutical product imparted with a 5α-reductase inhibitory function, and more specifically, an oriental pumpkin (Cucurbita moschata) and / or a western pumpkin (Cucurbita maxima). (5) A 5α-reductase inhibitor characterized by containing seeds as an active ingredient, and a food / beverage product and a pharmaceutical product imparted with a 5α-reductase inhibitory function.

  Testosterone (male hormone) biosynthesized and secreted in the testis and adrenal gland is transferred to the sebaceous glands, hair follicles, prostate, etc. via the blood, and then reduced by 5α-reductase, a metabolic enzyme present in each tissue. Converted to active dihydrotestosterone (DHT). Dihydrotestosterone (DHT) produced in each tissue is known to promote sebum secretion in the sebaceous glands and hair follicles and to promote cell proliferation in the prostate (see Non-Patent Document 1).

  At present, typical examples of diseases caused by excessive production of dihydrotestosterone by 5α-reductase include androgenetic alopecia, prostatic hypertrophy, acne vulgaris (acne) and the like. In general, synthetic drugs having a steroid structure are used as therapeutic agents for these diseases, but there are problems such as the appearance of undesirable hormone-like effects and dosage forms. However, at present, there are no therapeutic agents for the above-mentioned diseases that do not have a steroid structure.

Among compounds obtained by synthesis of compounds having no steroid structure isolated from mulberry or licorice, those having a 5α-reductase inhibitory action have been reported. (For example, refer to Patent Documents 1 and 2). However, since taking medicines obtained by chemical synthesis often involves side effects, taking them for a long time is not always desirable.
Therefore, a 5α-reductase inhibitor that is a component derived from a natural product and can be safely ingested over a long period of time without fear of side effects and rebounds, and a food and drink provided with a 5α-reductase inhibitory function and 5α-reductase inhibition Development of pharmaceuticals with added functions is desired.

  From such a viewpoint, attempts have been made to inhibit and improve the function of 5α-reductase in the body with food materials that can be administered on a daily basis. From natural products, extracts such as yacon (for example, refer to Patent Document 3) and spinach (for example, refer to Patent Document 4), which are native to Peru, are disclosed as testosterone 5α-reductase inhibitors.

However, these have problems of requiring a relatively large amount of administration, flavor problems, and storage stability problems such as precipitation during storage when blended in a beverage. And these plants receive a big restriction | limiting in terms of quantity when using as a raw material.
Japanese Unexamined Patent Publication No. 3-68518 Japanese Unexamined Patent Publication No. 7-17858 JP 2006-273755 A Japanese Patent No. 3276327 J.Steroid Biochemistry, 11,609 (1979)

  Conventional anti-androgenic hormones, especially steroids, have unfavorable side effects due to their strong hormonal action and have had problems with safety. Moreover, although the testosterone 5α-reductase inhibitors derived from natural products known so far have an inhibitory action, many of their active ingredients are unknown. Or even if the active ingredient was specified, there were many things with a weak effect | action.

  The present invention has been made in order to solve the above-mentioned problems, and the object of the present invention is to have a high safety that has an action of inhibiting excessive production of dihydrotestosterone by 5α-reductase and has no hormone-like action. An object of the present invention is to provide a 5α-reductase inhibitor, a food / beverage product and a pharmaceutical product imparted with a 5α-reductase inhibitory function. By inhibiting excessive production of dihydrotestosterone by 5α-reductase in vivo, it is useful for the prevention and treatment of diseases associated with 5α-reductase in vivo, especially prostatic hypertrophy, androgenetic alopecia, acne, etc. It is valid.

  In addition, ingesting food materials that can be administered on a daily basis in addition to medical treatment for various diseases and symptoms related to 5α-reductase can inhibit the action of 5α-reductase in vivo. Moreover, this invention makes it a subject to provide the food-drinks and pharmaceuticals which gave 5 alpha-reductase inhibitory function.

  The present inventor has conducted extensive research on the pharmacological action of pumpkin seeds, and remarkably inhibits overproduction of dihydrotestosterone by 5α-reductase in oriental pumpkin (Cucurbita moschata) and western pumpkin (Cucurbita maxima) seeds. It was found that there is an effect.

  Accordingly, the present invention is a 5α-reductase inhibitor characterized by containing oriental pumpkin (Cucurbita moschata) and / or western pumpkin (Cucurbita maxima) seeds as active ingredients. In addition, the present invention provides a food and drink provided with a 5α-reductase inhibitory function, comprising oriental pumpkin (Cucurbita moschata) and / or western pumpkin (Cucurbita maxima) seeds as an active ingredient, and an oriental pumpkin (Cucurbita moschata). And / or a pharmaceutical product imparted with a 5α-reductase inhibitory function, characterized by containing seeds of Cucurbita maxima as an active ingredient.

  The present invention provides a 5α-reductase inhibitor characterized by containing oriental pumpkin (Cucurbita moschata) and / or western pumpkin (Cucurbita maxima) seeds as an active ingredient, and also provides an oriental pumpkin (Cucurbita moschata). And / or food and drink with a 5α-reductase inhibitory function, characterized by containing western pumpkin (Cucurbita maxima) seeds as an active ingredient, and oriental pumpkin (Cucurbita moschata) and / or western pumpkin (Cucurbita maxima) seeds It was possible to solve the above-mentioned problems by providing a pharmaceutical product imparted with a 5α-reductase inhibitory function characterized by containing as an active ingredient.

  The 5α-reductase inhibitor of the present invention can be inhibited by ingesting the 5α-reductase inhibitory function, the food and drink provided with the 5α-reductase inhibitory function, and the pharmaceutical product provided with the 5α-reductase inhibitory function. Therefore, it is useful for prevention and improvement of diseases involving 5α-reductase.

  The 5α-reductase inhibitor according to the present invention is a food / beverage product imparted with a 5α-reductase inhibitory function, and a pharmaceutical product imparted with a 5α-reductase inhibitory function comprising oriental pumpkin (Cucurbita moschata) and / or western pumpkin (Cucurbita maxima) seeds. It contains as an active ingredient.

  Pumpkin is an annual or perennial plant of the cucurbitaceae, and is roughly divided into three types: oriental pumpkin (Cucurbita moschata), western pumpkin (Cucurbita maxima), pepo pumpkin (Cucurbita pepo). Or it is mainly a western pumpkin that is processed into frozen food or cooked food. Oriental pumpkins are said to be Japanese pumpkins and are sticky and sticky, but now Western pumpkins are almost consumed. Pepo pumpkins are very different from oriental and western pumpkins that have shelled seeds, and are characterized by having shellless seeds, which are rarely cultivated in Japan. There are small ones, elongated ones, and ones having a peculiar shape, and zucchini used in Western cuisine is a kind of peppo pumpkin, but peppo pumpkins other than zucchini are often used for decoration rather than edible. In addition, it is oil extracted from pepo pumpkin seeds that is produced overseas as edible oil (pumpkin seed oil). In the present invention, conventionally known fats and oils extracted from peppo pumpkin seeds are called pumpkin seed oils, and fats and oils extracted from western pumpkins or oriental pumpkin seeds tried in the present invention are called pumpkin seed oils.

  The present inventors have conducted extensive research on the pharmacological action of pumpkins, and found that the seeds of oriental pumpkins and western pumpkins have the effect of remarkably inhibiting the excessive production of dihydrotestosterone by 5α-reductase in vivo. I found.

  Regarding the seeds of pepo pumpkin, it is described in an electronic technical information advertisement when used for the treatment of urinary symptoms related to benign prostatic hypertrophy (CUCURBITA PEPO L. Pumpkin Seed Extract, Indena Japan Co., Ltd. Internet, <www.indena .com / pdf / cucurbita_pepo_jp.pdf>, search on January 26, 2007). However, the pumpkin described in this electronic technical information is clearly the seed of Cucurbita pepo, the Oriental pumpkin (Cucurbita moschata) and Western pumpkin (Cucurbita maxima) found by the present inventors Not a seed. As described above, various effects have been described under the names of “pumpkin seed” and “pumpkin seed oil”, but these are derived from peppo pumpkin and are currently generally used in Japan as edible. Or it is not a western pumpkin or an oriental pumpkin processed into frozen food or cooked food. Furthermore, it has not been known that oriental pumpkin seeds or western pumpkin seeds have an action of significantly inhibiting 5α-reductase in vivo.

  Oriental pumpkin (Cucurbita moschata) and western pumpkin (Cucurbita maxima) seeds, which are raw materials for the 5α-reductase inhibitor, etc. of the present invention, are agricultural wastes when processed into frozen foods. About 22,000 tons are discharged per year. Pumpkin seeds and cotton discharged during processing have been used as feed for livestock and composted to farmland, but due to distance problems with dairy farmers due to concentration of processing factories and intensive discharge. In addition, consumption as feed has reached its limit. It cannot be said that it is suitable for compost because it germinates for use as compost or it takes a long time to complete. For this reason, attempts to industrially make fertilizers and feeds have also been made, but high costs in the crushing and drying processes have become issues, making it difficult to put to practical use. Thus, oriental pumpkin (Cucurbita moschata) and western pumpkin (Cucurbita maxima) seeds are both unused biomass and waste. The present invention is also useful in terms of effective utilization of this unused biomass and reduction of waste.

  As shown in the following test examples, the action of significantly inhibiting 5α-reductase is 5α in the presence of commercially available pumpkin seed oil derived from oriental pumpkin seed oil, western pumpkin seed oil, and peppo pumpkin against an enzyme solution prepared from rat liver. -Reductase activity was measured. As a result, inhibition of 5α-reductase activity increased in proportion to the added concentrations of oriental pumpkin seed oil, western pumpkin seed oil and pumpkin seed oil. Therefore, it was clarified that a substance that inhibits the activity of 5α-reductase exists in the Oriental pumpkin seed oil and the Western pumpkin seed component. Moreover, it was found that the oriental pumpkin seed oil and the western pumpkin seed oil have significantly higher 5α-reductase inhibitory activity, which is significantly higher than the 5α-reductase inhibitory activity of the peppo pumpkin seed oil.

That is, the 5α-reductase inhibitor according to the present invention, the food and drink and the pharmaceutical product imparted with the 5α-reductase inhibitory function include oriental pumpkin or western pumpkin seeds.
The 5α-reductase inhibitor containing the oriental pumpkin or the western pumpkin seeds as an active ingredient according to the present invention, the food and drink and the pharmaceutical product imparted with the 5α-reductase inhibitory function, have an excellent 5α-reductase inhibitory action, so -Effective for prevention and treatment of prostatic hypertrophy, androgenetic alopecia, acne, etc., which are said to involve reductase.

  In the present invention, oriental pumpkin seeds or western pumpkin seeds are used as an active ingredient of a 5α-reductase inhibitor or to impart a 5α-reductase inhibitory function to foods and drinks and pharmaceuticals. As a kind of pumpkin that can be used in producing the 5α-reductase inhibitor as described above, as a result of the test example of the present invention, 5α-reductase inhibition significantly higher than the 5α-reductase inhibitory activity of peppo pumpkin seed oil. Oriental pumpkin (Cucurbita moschata) seeds and / or western pumpkin (Cucurbita maxima) seeds found to have activity are used.

The method for producing the 5α-reductase inhibitor of the present invention is roughly as follows.
Extract lipids from seeds of oriental pumpkin or western pumpkin. Extraction of lipids,
After the seeds pressurized heat, rather good in heat pressing method to collect squeezing the oil toward expeller, oil component collected is purified except After filtering off the precipitate formed by adding water by centrifugation. Also, pressurized thermal pressing method has poor extraction efficiency of oil extraction residue which remains considerable oil content may be to use a pressure抽法you combination solvent process for the remaining oil recovery. The pressure extraction method extracts oil remaining hexane was added to the extraction residue of the pressurized heat pressing method, oil of evaporated is effective in terms of efficiency because the purification together with oil heating pressing method.

  The oriental pumpkin or western pumpkin seed oil refined using any of the above extraction methods exhibits a bright yellow transparent liquid, and is used as it is or in combination with other raw materials as the 5α-reductase inhibitor of the present invention. It can be used as an active ingredient or as a food or drink or pharmaceutical product having a 5α-reductase inhibitory function.

  In addition, the dosage form of the 5α-reductase inhibitor and the pharmaceutical agent imparted with the 5α-reductase inhibitory function of the present invention is not particularly limited. For example, for treatment of benign prostatic hyperplasia, powders, tablets, capsules , Granules, powders, liquids, etc., orally. Moreover, these dosage forms can be manufactured by the conventional formulation method known conventionally. For example, it is mixed with a carrier or excipient that is permitted in the production of the preparation and molded. Examples of excipients include lactose, dextrose, cellulose, methylcellulose, starch and the like.

  In addition, for the 5α-reductase-inhibiting food or drink of the present invention, purified oriental pumpkin or western pumpkin seed oil is used as a food or drink of the above-mentioned type, or purified oriental pumpkin or western pumpkin seed oil is consumed or consumed. Blend into the product.

  The dose of the 5α-reductase-inhibiting lipid metabolism-improving agent, the 5α-reductase-inhibiting lipid metabolism-improving food and drink and the pharmaceutical product imparted with the 5α-reductase-inhibiting function of the present invention varies depending on age, therapeutic effect, disease state, etc., and is not particularly limited. However, the blending amount and the like may be adjusted so that the refined oriental pumpkin seed oil or western pumpkin seed oil can be ingested in an amount of 0.1 to 500 mg, preferably 0.5 to 100 mg.

The following Example illustrates the reference Rei及 beauty Test Examples, but the present invention will be described in detail, these are merely illustrative, the present invention is not limited thereto by any means.

[Reference Example 1]
(Preparation of Western pumpkin seed oil 1)
After carefully grinding the selected western pumpkin seeds, the oil content was squeezed and collected using a press. The oil was separated by filtration, and the precipitate formed by adding water was removed by centrifugation, followed by purification to obtain western pumpkin seed oil. Yield was 25% .

(Preparation of Western pumpkin seed oil 2)
Hits were squash seeds Water was added to the heated pressurized, collected squeezing oil subjected mill. The oil was separated by filtration, and the precipitate formed by adding water was removed by centrifugation, followed by purification to obtain western pumpkin seed oil. Yield was 50%. This can be used as it is or as a 5α-reductase inhibitor of the present invention by mixing it with other raw materials and filling it into a container such as a bottle, can or plastic. Further, by formulating into various dosage forms, you can use as a medicament which endow 5α- reductase inhibitor and 5α- reductase inhibition function.

(Preparation of Western pumpkin seed oil 3)
Hits were squash seeds Water was added to the heated pressurized, collected squeezing oil subjected mill. Hexane was added to this extraction residue to extract the remaining oil, and the oil obtained by distilling off the solvent was mixed with the oil obtained by the above-mentioned pressing and purified to obtain Western pumpkin seed oil. Yield was 80%. This can be used as it is or as a 5α-reductase inhibitor of the present invention by mixing it with other raw materials and filling it into a container such as a bottle, can or plastic. Further, by formulating into various dosage forms, you can use as a medicament which endow 5α- reductase inhibitor and 5α- reductase inhibition function.

[Reference Example 2]
(Preparation of oriental pumpkin seed oil 1)
An oriental pumpkin seed oil was obtained under the same conditions as in Reference Example 1 except that a carefully selected oriental pumpkin seed was used .

(Preparation of oriental pumpkin seed oil 2)
An oriental pumpkin seed oil was obtained under the same conditions as in Example 1 except that the carefully selected oriental pumpkin seeds were used. This can be used as it is or as a 5α-reductase inhibitor of the present invention by mixing it with other raw materials and filling it into a container such as a bottle, can or plastic. Further, by formulating into various dosage forms, you can use as a medicament which endow 5α- reductase inhibitor and 5α- reductase inhibition function.

(Preparation of oriental pumpkin seed oil 3)
An oriental pumpkin seed oil was obtained under the same conditions as in Example 2 except that the carefully selected oriental pumpkin seeds were used. This can be used as it is or as a 5α-reductase inhibitor of the present invention by being filled with a container such as a bottle, can, or plastic after blending with other raw materials. Further, by formulating into various dosage forms, you can use as a medicament which endow 5α- reductase inhibitor and 5α- reductase inhibition function.

[Test Example 1]
Using the Western pumpkin seed oil obtained in Example 1 , the Oriental pumpkin seed oil obtained in Example 3 and the commercially available peppo pumpkin seed oil as samples, an enzyme solution prepared from the liver of the rat was used to inhibit 5α-reductase. Examined.
1) Preparation of enzyme solution Eight-week-old male SD rats were fasted for 24 hours and then laparotomized to extract the liver. The extracted liver was perfused with Krebs-Ringer phosphate buffer (pH 7.4) to remove blood and freeze-fixed. The frozen and fixed liver was pulverized in a mortar and then homogenated by adding 5 volumes of 10 mmol / L Tris-HCl buffer (pH 7.2). This was centrifuged at 9,000 × g for 10 minutes at 4 ° C., and the supernatant was collected and stored frozen at −80 ° C.

2) Preparation of reaction solution A reaction solution having the composition shown in Table 1 was prepared. The western pumpkin seed oil obtained in Example 1 was used as the western pumpkin seed oil, and the oriental pumpkin seed oil obtained in Example 3 was used as the oriental pumpkin seed oil. A commercially available pumpkin seed oil (Wolf) was used as the peppo pumpkin seed oil. Sample A to which no enzyme solution was added was designated as Sample A. Sample B was prepared by adding the enzyme solution and not adding pepo pumpkin seed oil, western pumpkin seed oil or oriental pumpkin seed oil. Sample C was prepared by adding an enzyme solution and adding peppo pumpkin seed oil, western pumpkin seed oil, or oriental pumpkin seed oil. As the testosterone solution, 500 μg / ml (propylene glycol: 10 mM Tris-hydrochloric acid buffer (pH 7.2) = 1: 1 v / v) was used. As the coenzyme solution, NADPH (nicotinamide adenine dinucleotide phosphate) was used. The buffer used was 10 mM Tris-hydrochloric acid buffer (pH 7.2). The test substance, western pumpkin seed oil, oriental pumpkin seed oil, or peppo pumpkin seed oil, was mixed with a testosterone solution, a buffer solution was added thereto, and an enzyme solution and a coenzyme solution were added and reacted.

  In addition, Table 2 shows the composition of the western pumpkin seed oil, oriental pumpkin seed oil, or peppo pumpkin seed oil addition solution. The seed oil used was a 50% ethanol solution, and the buffer used was 10 mM Tris-hydrochloric acid buffer (pH 7.2).

3) Measurement pretreatment The reaction solution was incubated at 37 ° C. for 30 minutes, 0.05 ml of 70% perchloric acid was added, and the solution was added to a solid phase extraction column (OASIS HLB). 2 ml of purified water was passed through the column for washing, and then 2 ml of 40% methanol was passed through the column for washing. 1 ml of eluate (100% methanol) was passed through the column to elute testosterone, and the eluate was evaporated to dryness under reduced pressure. The residue was dissolved with 0.25 ml of 65% methanol. 0.01 ml of this lysate was used for measurement by high performance liquid chromatography (HPLC).

4) HPLC conditions The column was YMC-Pak ODS-AM, and the measurement was performed at a mobile phase of methanol: water = 65: 35 (v / v) at around 40 ° C. and at a flow rate of 1.0 ml / min. For detection, an ultraviolet absorptiometer (measurement wavelength: 254 nm) was used.

5) Inhibition rate calculation formula The inhibition rate was calculated by the following formula.

Inhibition rate (%) = 100− (sample A peak area−sample C peak area) / (sample A peak area−sample B peak area) × 100

The peak area represents the peak area of testosterone in the HPLC chromatogram.

6) The 5α-reductase inhibitory action was examined. The solid phase extraction method was used as a testosterone extraction method in the reaction solution, and the 5α-reductase activity in the presence of western pumpkin seed oil, oriental pumpkin seed oil, or peppo pumpkin seed oil was measured. The result is shown in FIG.
As shown in FIG. 1, the 5α-reductase inhibitory activity increased in proportion to the added concentration of western pumpkin seed oil, oriental pumpkin seed oil, or pepo pumpkin seed oil. And it was shown that Oriental pumpkin seed oil and Western pumpkin seed oil have 5α-reductase inhibitory activity which was not known conventionally. Moreover, it was found that the oriental pumpkin seed oil and the western pumpkin seed oil have significantly higher 5α-reductase inhibitory activity, which is significantly higher than the 5α-reductase inhibitory activity of the peppo pumpkin seed oil.

  From the above results, it was clarified that oriental pumpkin seeds and western pumpkin seeds have the effect of significantly inhibiting 5α-reductase. This makes it possible to use the western pumpkin or oriental pumpkin seed, which is a residue used for processing in the past and has been discarded, for a new and useful application of a 5α-reductase inhibitor.

5 shows 5α-reductase activity in the presence of pepo pumpkin seed oil, oriental pumpkin seed oil and western pumpkin seed oil (Test Example 1).

Claims (3)

A 5α-reductase inhibitor characterized by containing, as an active ingredient, an oil component obtained by heating seeds of oriental pumpkin (Cucurbita moschata) and / or western pumpkin (Cucurbita maxima) and squeezing them by a press . 5. The 5α-reductase inhibitor according to claim 1, which is an oil or fat obtained by heating seeds of oriental pumpkin (Cucurbita moschata) and / or western pumpkin (Cucurbita maxima) and squeezing them through a press . A medicinal product having a 5α-reductase inhibitory function, characterized by containing as an active ingredient an oil component obtained by heating seeds of oriental pumpkin (Cucurbita moschata) and / or western pumpkin (Cucurbita maxima) and squeezing them by a press .